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        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

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        About MultiQC

        This report was generated using MultiQC, version 1.9

        You can see a YouTube video describing how to use MultiQC reports here: https://youtu.be/qPbIlO_KWN0

        For more information about MultiQC, including other videos and extensive documentation, please visit http://multiqc.info

        You can report bugs, suggest improvements and find the source code for MultiQC on GitHub: https://github.com/ewels/MultiQC

        MultiQC is published in Bioinformatics:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

        A modular tool to aggregate results from bioinformatics analyses across many samples into a single report.

        Report generated on 2021-08-23, 11:45 based on data in: /scratch/gencore/logs/html/HGVMYDRXY/merged


        General Statistics

        Showing 118/118 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HGVMYDRXY_n01_CIVRCoV_P1_A01_C001
        75.1%
        56%
        30.0
        HGVMYDRXY_n01_CIVRCoV_P1_A02_U277
        69.6%
        57%
        32.0
        HGVMYDRXY_n01_CIVRCoV_P1_A03_C028
        90.3%
        59%
        32.9
        HGVMYDRXY_n01_CIVRCoV_P1_A04_U514
        77.6%
        57%
        30.3
        HGVMYDRXY_n01_CIVRCoV_P1_B01_C005
        84.9%
        59%
        26.0
        HGVMYDRXY_n01_CIVRCoV_P1_B02_C014
        82.0%
        59%
        21.1
        HGVMYDRXY_n01_CIVRCoV_P1_B03_U409
        82.0%
        59%
        104.5
        HGVMYDRXY_n01_CIVRCoV_P1_B04_C023
        83.4%
        58%
        30.1
        HGVMYDRXY_n01_CIVRCoV_P1_C01_U051
        79.8%
        58%
        30.9
        HGVMYDRXY_n01_CIVRCoV_P1_C02_U288
        77.7%
        58%
        27.7
        HGVMYDRXY_n01_CIVRCoV_P1_C03_C011
        57.5%
        54%
        22.1
        HGVMYDRXY_n01_CIVRCoV_P1_C04_U506
        67.0%
        56%
        26.3
        HGVMYDRXY_n01_CIVRCoV_P1_D01_C007
        87.4%
        59%
        27.8
        HGVMYDRXY_n01_CIVRCoV_P1_D02_U062
        70.1%
        57%
        19.8
        HGVMYDRXY_n01_CIVRCoV_P1_D03_U396
        63.8%
        55%
        28.1
        HGVMYDRXY_n01_CIVRCoV_P1_D04_C019
        66.7%
        55%
        29.3
        HGVMYDRXY_n01_CIVRCoV_P1_E01_U175
        64.1%
        56%
        24.4
        HGVMYDRXY_n01_CIVRCoV_P1_E02_U327
        65.6%
        56%
        28.2
        HGVMYDRXY_n01_CIVRCoV_P1_E03_C026
        69.3%
        57%
        14.0
        HGVMYDRXY_n01_CIVRCoV_P1_E04_Neg1
        53.6%
        62%
        0.0
        HGVMYDRXY_n01_CIVRCoV_P1_F01_C004
        88.7%
        59%
        35.2
        HGVMYDRXY_n01_CIVRCoV_P1_F02_C025
        84.4%
        59%
        30.6
        HGVMYDRXY_n01_CIVRCoV_P1_F03_U470
        73.9%
        58%
        17.6
        HGVMYDRXY_n01_CIVRCoV_P1_G01_C008
        67.0%
        55%
        16.3
        HGVMYDRXY_n01_CIVRCoV_P1_G02_U264
        70.5%
        56%
        26.1
        HGVMYDRXY_n01_CIVRCoV_P1_G03_C015
        86.5%
        59%
        30.9
        HGVMYDRXY_n01_CIVRCoV_P1_H01_U055
        74.8%
        57%
        30.5
        HGVMYDRXY_n01_CIVRCoV_P1_H02_C020
        77.7%
        58%
        30.5
        HGVMYDRXY_n01_CIVRCoV_P1_H03_C021
        70.5%
        57%
        25.3
        HGVMYDRXY_n01_CIVRCoV_P2_A05_U420
        61.8%
        55%
        30.2
        HGVMYDRXY_n01_CIVRCoV_P2_A06_C012
        80.8%
        57%
        29.0
        HGVMYDRXY_n01_CIVRCoV_P2_A07_C027
        78.4%
        58%
        27.7
        HGVMYDRXY_n01_CIVRCoV_P2_A08_U340
        55.9%
        52%
        32.9
        HGVMYDRXY_n01_CIVRCoV_P2_B05_C010
        74.5%
        57%
        28.2
        HGVMYDRXY_n01_CIVRCoV_P2_B06_U467
        67.3%
        57%
        30.9
        HGVMYDRXY_n01_CIVRCoV_P2_B07_U477
        61.8%
        56%
        28.6
        HGVMYDRXY_n01_CIVRCoV_P2_B08_Neg2
        29.9%
        55%
        0.0
        HGVMYDRXY_n01_CIVRCoV_P2_C05_U008
        78.7%
        57%
        35.1
        HGVMYDRXY_n01_CIVRCoV_P2_C06_C024
        71.5%
        58%
        31.7
        HGVMYDRXY_n01_CIVRCoV_P2_C07_C006
        64.7%
        54%
        36.9
        HGVMYDRXY_n01_CIVRCoV_P2_D05_U194
        72.6%
        58%
        33.3
        HGVMYDRXY_n01_CIVRCoV_P2_D06_U487
        51.4%
        53%
        27.9
        HGVMYDRXY_n01_CIVRCoV_P2_D07_U439
        66.6%
        55%
        30.0
        HGVMYDRXY_n01_CIVRCoV_P2_E05_C002
        79.2%
        58%
        32.7
        HGVMYDRXY_n01_CIVRCoV_P2_E06_U356
        74.1%
        57%
        26.7
        HGVMYDRXY_n01_CIVRCoV_P2_E07_U244
        75.0%
        58%
        43.9
        HGVMYDRXY_n01_CIVRCoV_P2_F04_C003
        79.4%
        58%
        33.1
        HGVMYDRXY_n01_CIVRCoV_P2_F05_U488
        55.6%
        54%
        31.3
        HGVMYDRXY_n01_CIVRCoV_P2_F06_C009
        71.3%
        56%
        36.2
        HGVMYDRXY_n01_CIVRCoV_P2_F07_U030
        68.9%
        57%
        37.7
        HGVMYDRXY_n01_CIVRCoV_P2_G04_C018
        77.7%
        57%
        30.4
        HGVMYDRXY_n01_CIVRCoV_P2_G05_U128
        71.0%
        56%
        38.6
        HGVMYDRXY_n01_CIVRCoV_P2_G06_C022
        77.1%
        56%
        30.4
        HGVMYDRXY_n01_CIVRCoV_P2_G07_C017
        80.6%
        59%
        38.2
        HGVMYDRXY_n01_CIVRCoV_P2_H04_U236
        70.6%
        57%
        31.3
        HGVMYDRXY_n01_CIVRCoV_P2_H05_C013
        70.2%
        56%
        34.4
        HGVMYDRXY_n01_CIVRCoV_P2_H06_U482
        77.4%
        58%
        27.6
        HGVMYDRXY_n01_CIVRCoV_P2_H07_C016
        69.5%
        56%
        29.2
        HGVMYDRXY_n01_undetermined
        62.0%
        52%
        131.0
        HGVMYDRXY_n02_CIVRCoV_P1_A01_C001
        74.1%
        56%
        30.0
        HGVMYDRXY_n02_CIVRCoV_P1_A02_U277
        67.8%
        57%
        32.0
        HGVMYDRXY_n02_CIVRCoV_P1_A03_C028
        89.1%
        59%
        32.9
        HGVMYDRXY_n02_CIVRCoV_P1_A04_U514
        76.1%
        57%
        30.3
        HGVMYDRXY_n02_CIVRCoV_P1_B01_C005
        83.0%
        59%
        26.0
        HGVMYDRXY_n02_CIVRCoV_P1_B02_C014
        80.4%
        59%
        21.1
        HGVMYDRXY_n02_CIVRCoV_P1_B03_U409
        80.2%
        58%
        104.5
        HGVMYDRXY_n02_CIVRCoV_P1_B04_C023
        82.1%
        58%
        30.1
        HGVMYDRXY_n02_CIVRCoV_P1_C01_U051
        77.7%
        58%
        30.9
        HGVMYDRXY_n02_CIVRCoV_P1_C02_U288
        76.1%
        58%
        27.7
        HGVMYDRXY_n02_CIVRCoV_P1_C03_C011
        56.9%
        54%
        22.1
        HGVMYDRXY_n02_CIVRCoV_P1_C04_U506
        66.0%
        56%
        26.3
        HGVMYDRXY_n02_CIVRCoV_P1_D01_C007
        86.0%
        59%
        27.8
        HGVMYDRXY_n02_CIVRCoV_P1_D02_U062
        68.5%
        57%
        19.8
        HGVMYDRXY_n02_CIVRCoV_P1_D03_U396
        62.6%
        55%
        28.1
        HGVMYDRXY_n02_CIVRCoV_P1_D04_C019
        65.7%
        55%
        29.3
        HGVMYDRXY_n02_CIVRCoV_P1_E01_U175
        62.9%
        56%
        24.4
        HGVMYDRXY_n02_CIVRCoV_P1_E02_U327
        63.5%
        56%
        28.2
        HGVMYDRXY_n02_CIVRCoV_P1_E03_C026
        68.0%
        57%
        14.0
        HGVMYDRXY_n02_CIVRCoV_P1_E04_Neg1
        26.7%
        63%
        0.0
        HGVMYDRXY_n02_CIVRCoV_P1_F01_C004
        87.3%
        59%
        35.2
        HGVMYDRXY_n02_CIVRCoV_P1_F02_C025
        82.9%
        59%
        30.6
        HGVMYDRXY_n02_CIVRCoV_P1_F03_U470
        72.2%
        58%
        17.6
        HGVMYDRXY_n02_CIVRCoV_P1_G01_C008
        65.5%
        55%
        16.3
        HGVMYDRXY_n02_CIVRCoV_P1_G02_U264
        69.2%
        56%
        26.1
        HGVMYDRXY_n02_CIVRCoV_P1_G03_C015
        84.6%
        59%
        30.9
        HGVMYDRXY_n02_CIVRCoV_P1_H01_U055
        73.3%
        57%
        30.5
        HGVMYDRXY_n02_CIVRCoV_P1_H02_C020
        76.0%
        58%
        30.5
        HGVMYDRXY_n02_CIVRCoV_P1_H03_C021
        68.9%
        57%
        25.3
        HGVMYDRXY_n02_CIVRCoV_P2_A05_U420
        60.1%
        55%
        30.2
        HGVMYDRXY_n02_CIVRCoV_P2_A06_C012
        79.3%
        57%
        29.0
        HGVMYDRXY_n02_CIVRCoV_P2_A07_C027
        76.9%
        58%
        27.7
        HGVMYDRXY_n02_CIVRCoV_P2_A08_U340
        54.4%
        52%
        32.9
        HGVMYDRXY_n02_CIVRCoV_P2_B05_C010
        73.1%
        57%
        28.2
        HGVMYDRXY_n02_CIVRCoV_P2_B06_U467
        65.8%
        56%
        30.9
        HGVMYDRXY_n02_CIVRCoV_P2_B07_U477
        60.4%
        56%
        28.6
        HGVMYDRXY_n02_CIVRCoV_P2_B08_Neg2
        13.8%
        56%
        0.0
        HGVMYDRXY_n02_CIVRCoV_P2_C05_U008
        77.3%
        57%
        35.1
        HGVMYDRXY_n02_CIVRCoV_P2_C06_C024
        70.4%
        58%
        31.7
        HGVMYDRXY_n02_CIVRCoV_P2_C07_C006
        63.4%
        54%
        36.9
        HGVMYDRXY_n02_CIVRCoV_P2_D05_U194
        70.3%
        58%
        33.3
        HGVMYDRXY_n02_CIVRCoV_P2_D06_U487
        49.8%
        53%
        27.9
        HGVMYDRXY_n02_CIVRCoV_P2_D07_U439
        65.5%
        55%
        30.0
        HGVMYDRXY_n02_CIVRCoV_P2_E05_C002
        77.5%
        57%
        32.7
        HGVMYDRXY_n02_CIVRCoV_P2_E06_U356
        72.3%
        57%
        26.7
        HGVMYDRXY_n02_CIVRCoV_P2_E07_U244
        73.9%
        58%
        43.9
        HGVMYDRXY_n02_CIVRCoV_P2_F04_C003
        77.9%
        58%
        33.1
        HGVMYDRXY_n02_CIVRCoV_P2_F05_U488
        54.6%
        54%
        31.3
        HGVMYDRXY_n02_CIVRCoV_P2_F06_C009
        70.1%
        55%
        36.2
        HGVMYDRXY_n02_CIVRCoV_P2_F07_U030
        67.6%
        57%
        37.7
        HGVMYDRXY_n02_CIVRCoV_P2_G04_C018
        76.3%
        57%
        30.4
        HGVMYDRXY_n02_CIVRCoV_P2_G05_U128
        69.6%
        56%
        38.6
        HGVMYDRXY_n02_CIVRCoV_P2_G06_C022
        75.5%
        56%
        30.4
        HGVMYDRXY_n02_CIVRCoV_P2_G07_C017
        79.1%
        59%
        38.2
        HGVMYDRXY_n02_CIVRCoV_P2_H04_U236
        69.1%
        57%
        31.3
        HGVMYDRXY_n02_CIVRCoV_P2_H05_C013
        69.0%
        56%
        34.4
        HGVMYDRXY_n02_CIVRCoV_P2_H06_U482
        75.8%
        58%
        27.6
        HGVMYDRXY_n02_CIVRCoV_P2_H07_C016
        67.7%
        56%
        29.2
        HGVMYDRXY_n02_undetermined
        52.5%
        53%
        131.0

        Demultiplexing Report


        Total Read Count: Total number of PF (Passing Filter) reads in this library.
        Portion: The proportion of reads that represent the individual library in the entire Library Pool.

        Showing 59/59 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        131027373
        7.0
        CIVRCoV_P1_A01_C001
        29956518
        1.6
        CIVRCoV_P1_B01_C005
        25969534
        1.4
        CIVRCoV_P1_C01_U051
        30911873
        1.7
        CIVRCoV_P1_D01_C007
        27779669
        1.5
        CIVRCoV_P1_E01_U175
        24376890
        1.3
        CIVRCoV_P1_F01_C004
        35195663
        1.9
        CIVRCoV_P1_G01_C008
        16340167
        0.9
        CIVRCoV_P1_H01_U055
        30473094
        1.6
        CIVRCoV_P1_A02_U277
        32047296
        1.7
        CIVRCoV_P1_B02_C014
        21095587
        1.1
        CIVRCoV_P1_C02_U288
        27693367
        1.5
        CIVRCoV_P1_D02_U062
        19764564
        1.1
        CIVRCoV_P1_E02_U327
        28221049
        1.5
        CIVRCoV_P1_F02_C025
        30569173
        1.6
        CIVRCoV_P1_G02_U264
        26126472
        1.4
        CIVRCoV_P1_H02_C020
        30504607
        1.6
        CIVRCoV_P1_A03_C028
        32929992
        1.8
        CIVRCoV_P1_B03_U409
        104488271
        5.6
        CIVRCoV_P1_C03_C011
        22118081
        1.2
        CIVRCoV_P1_D03_U396
        28127469
        1.5
        CIVRCoV_P1_E03_C026
        14044623
        0.8
        CIVRCoV_P1_F03_U470
        17584008
        0.9
        CIVRCoV_P1_G03_C015
        30879529
        1.7
        CIVRCoV_P1_H03_C021
        25314101
        1.4
        CIVRCoV_P1_A04_U514
        30290668
        1.6
        CIVRCoV_P1_B04_C023
        30144415
        1.6
        CIVRCoV_P1_C04_U506
        26313908
        1.4
        CIVRCoV_P1_D04_C019
        29341634
        1.6
        CIVRCoV_P1_E04_Neg1
        21165
        0.0
        CIVRCoV_P2_F04_C003
        33139787
        1.8
        CIVRCoV_P2_G04_C018
        30423997
        1.6
        CIVRCoV_P2_H04_U236
        31319575
        1.7
        CIVRCoV_P2_A05_U420
        30172153
        1.6
        CIVRCoV_P2_B05_C010
        28246254
        1.5
        CIVRCoV_P2_C05_U008
        35085553
        1.9
        CIVRCoV_P2_D05_U194
        33328622
        1.8
        CIVRCoV_P2_E05_C002
        32696149
        1.8
        CIVRCoV_P2_F05_U488
        31346907
        1.7
        CIVRCoV_P2_G05_U128
        38589110
        2.1
        CIVRCoV_P2_H05_C013
        34408420
        1.8
        CIVRCoV_P2_A06_C012
        28999623
        1.6
        CIVRCoV_P2_B06_U467
        30877801
        1.7
        CIVRCoV_P2_C06_C024
        31704453
        1.7
        CIVRCoV_P2_D06_U487
        27917483
        1.5
        CIVRCoV_P2_E06_U356
        26747068
        1.4
        CIVRCoV_P2_F06_C009
        36230881
        1.9
        CIVRCoV_P2_G06_C022
        30440733
        1.6
        CIVRCoV_P2_H06_U482
        27619818
        1.5
        CIVRCoV_P2_A07_C027
        27737187
        1.5
        CIVRCoV_P2_B07_U477
        28602196
        1.5
        CIVRCoV_P2_C07_C006
        36936115
        2.0
        CIVRCoV_P2_D07_U439
        29998806
        1.6
        CIVRCoV_P2_E07_U244
        43883354
        2.4
        CIVRCoV_P2_F07_U030
        37711504
        2.0
        CIVRCoV_P2_G07_C017
        38239736
        2.1
        CIVRCoV_P2_H07_C016
        29152133
        1.6
        CIVRCoV_P2_A08_U340
        32910639
        1.8
        CIVRCoV_P2_B08_Neg2
        28201
        0.0

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        2553348096
        1864145018
        7.0
        0.7

        Barcodes of Undetermined Reads


        We have determined the barcodes of your undetermined reads (reads containing a barcode that you did not encode in your metadata). Here are the top 20 barcodes belonging to the undetermined reads. The full list is available here.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGGGGAGATCTCG
        8673938.0
        6.6
        GGGGGGGGCGATCTCG
        3292807.0
        2.5
        GTACCACAGGGTGTGG
        2344310.0
        1.8
        CTGTACCACGGCTGTG
        1750848.0
        1.3
        GCGTTAGAGGCTGCTG
        1011676.0
        0.8
        GTACCACAGGGGGTGG
        894541.0
        0.7
        GGTATAGGGGGGAGTA
        873998.0
        0.7
        ATCTGACCCCGTCGAG
        754237.0
        0.6
        GGAGGAATCCCCGTTC
        713258.0
        0.5
        GTACCACAGGGGGGGG
        703625.0
        0.5
        ATAACGCCGGACCCTG
        699362.0
        0.5
        GTCCTAAGGGGGAGCT
        640277.0
        0.5
        CTGTACCACGTCGGTG
        613740.0
        0.5
        GTCCTAAGGGGTCGCT
        597361.0
        0.5
        TTACCGACGGGATTCG
        589500.0
        0.5
        TACCTGCAGGCTGTCC
        573133.0
        0.4
        TACCTGCAGGTTGTCC
        564250.0
        0.4
        GTACCACAGGGTGGGG
        563958.0
        0.4
        CCTTCCATCGTGCGTG
        529132.0
        0.4
        GTACCACAGGTGGTGG
        525963.0
        0.4

        FastQC

        FastQC is a quality control tool for high throughput sequence data, written by Simon Andrews at the Babraham Institute in Cambridge.

        Sequence Counts

        Sequence counts for each sample. Duplicate read counts are an estimate only.

        This plot show the total number of reads, broken down into unique and duplicate if possible (only more recent versions of FastQC give duplicate info).

        You can read more about duplicate calculation in the FastQC documentation. A small part has been copied here for convenience:

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Quality Histograms

        The mean quality value across each base position in the read.

        To enable multiple samples to be plotted on the same graph, only the mean quality scores are plotted (unlike the box plots seen in FastQC reports).

        Taken from the FastQC help:

        The y-axis on the graph shows the quality scores. The higher the score, the better the base call. The background of the graph divides the y axis into very good quality calls (green), calls of reasonable quality (orange), and calls of poor quality (red). The quality of calls on most platforms will degrade as the run progresses, so it is common to see base calls falling into the orange area towards the end of a read.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Sequence Quality Scores

        The number of reads with average quality scores. Shows if a subset of reads has poor quality.

        From the FastQC help:

        The per sequence quality score report allows you to see if a subset of your sequences have universally low quality values. It is often the case that a subset of sequences will have universally poor quality, however these should represent only a small percentage of the total sequences.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Base Sequence Content

        The proportion of each base position for which each of the four normal DNA bases has been called.

        To enable multiple samples to be shown in a single plot, the base composition data is shown as a heatmap. The colours represent the balance between the four bases: an even distribution should give an even muddy brown colour. Hover over the plot to see the percentage of the four bases under the cursor.

        To see the data as a line plot, as in the original FastQC graph, click on a sample track.

        From the FastQC help:

        Per Base Sequence Content plots out the proportion of each base position in a file for which each of the four normal DNA bases has been called.

        In a random library you would expect that there would be little to no difference between the different bases of a sequence run, so the lines in this plot should run parallel with each other. The relative amount of each base should reflect the overall amount of these bases in your genome, but in any case they should not be hugely imbalanced from each other.

        It's worth noting that some types of library will always produce biased sequence composition, normally at the start of the read. Libraries produced by priming using random hexamers (including nearly all RNA-Seq libraries) and those which were fragmented using transposases inherit an intrinsic bias in the positions at which reads start. This bias does not concern an absolute sequence, but instead provides enrichement of a number of different K-mers at the 5' end of the reads. Whilst this is a true technical bias, it isn't something which can be corrected by trimming and in most cases doesn't seem to adversely affect the downstream analysis.

        Click a sample row to see a line plot for that dataset.
        Rollover for sample name
        Position: -
        %T: -
        %C: -
        %A: -
        %G: -

        Per Sequence GC Content

        The average GC content of reads. Normal random library typically have a roughly normal distribution of GC content.

        From the FastQC help:

        This module measures the GC content across the whole length of each sequence in a file and compares it to a modelled normal distribution of GC content.

        In a normal random library you would expect to see a roughly normal distribution of GC content where the central peak corresponds to the overall GC content of the underlying genome. Since we don't know the the GC content of the genome the modal GC content is calculated from the observed data and used to build a reference distribution.

        An unusually shaped distribution could indicate a contaminated library or some other kinds of biased subset. A normal distribution which is shifted indicates some systematic bias which is independent of base position. If there is a systematic bias which creates a shifted normal distribution then this won't be flagged as an error by the module since it doesn't know what your genome's GC content should be.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Base N Content

        The percentage of base calls at each position for which an N was called.

        From the FastQC help:

        If a sequencer is unable to make a base call with sufficient confidence then it will normally substitute an N rather than a conventional base call. This graph shows the percentage of base calls at each position for which an N was called.

        It's not unusual to see a very low proportion of Ns appearing in a sequence, especially nearer the end of a sequence. However, if this proportion rises above a few percent it suggests that the analysis pipeline was unable to interpret the data well enough to make valid base calls.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Length Distribution

        All samples have sequences of a single length (151bp).

        Sequence Duplication Levels

        The relative level of duplication found for every sequence.

        From the FastQC Help:

        In a diverse library most sequences will occur only once in the final set. A low level of duplication may indicate a very high level of coverage of the target sequence, but a high level of duplication is more likely to indicate some kind of enrichment bias (eg PCR over amplification). This graph shows the degree of duplication for every sequence in a library: the relative number of sequences with different degrees of duplication.

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        In a properly diverse library most sequences should fall into the far left of the plot in both the red and blue lines. A general level of enrichment, indicating broad oversequencing in the library will tend to flatten the lines, lowering the low end and generally raising other categories. More specific enrichments of subsets, or the presence of low complexity contaminants will tend to produce spikes towards the right of the plot.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Overrepresented sequences

        The total amount of overrepresented sequences found in each library.

        FastQC calculates and lists overrepresented sequences in FastQ files. It would not be possible to show this for all samples in a MultiQC report, so instead this plot shows the number of sequences categorized as over represented.

        Sometimes, a single sequence may account for a large number of reads in a dataset. To show this, the bars are split into two: the first shows the overrepresented reads that come from the single most common sequence. The second shows the total count from all remaining overrepresented sequences.

        From the FastQC Help:

        A normal high-throughput library will contain a diverse set of sequences, with no individual sequence making up a tiny fraction of the whole. Finding that a single sequence is very overrepresented in the set either means that it is highly biologically significant, or indicates that the library is contaminated, or not as diverse as you expected.

        FastQC lists all of the sequences which make up more than 0.1% of the total. To conserve memory only sequences which appear in the first 100,000 sequences are tracked to the end of the file. It is therefore possible that a sequence which is overrepresented but doesn't appear at the start of the file for some reason could be missed by this module.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Adapter Content

        The cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position.

        Note that only samples with ≥ 0.1% adapter contamination are shown.

        There may be several lines per sample, as one is shown for each adapter detected in the file.

        From the FastQC Help:

        The plot shows a cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position. Once a sequence has been seen in a read it is counted as being present right through to the end of the read so the percentages you see will only increase as the read length goes on.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Status Checks

        Status for each FastQC section showing whether results seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        FastQC assigns a status for each section of the report. These give a quick evaluation of whether the results of the analysis seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        It is important to stress that although the analysis results appear to give a pass/fail result, these evaluations must be taken in the context of what you expect from your library. A 'normal' sample as far as FastQC is concerned is random and diverse. Some experiments may be expected to produce libraries which are biased in particular ways. You should treat the summary evaluations therefore as pointers to where you should concentrate your attention and understand why your library may not look random and diverse.

        Specific guidance on how to interpret the output of each module can be found in the relevant report section, or in the FastQC help.

        In this heatmap, we summarise all of these into a single heatmap for a quick overview. Note that not all FastQC sections have plots in MultiQC reports, but all status checks are shown in this heatmap.

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